SBIR Phase I: Recombinant Multiepitope Mosaic Protein Design for Urine-based Diagnosis of Leptospirosis

SBIR 第一阶段：用于钩端螺旋体病尿液诊断的重组多表位镶嵌蛋白设计

• 批准号: 1215242
• 负责人: Edwin Agbo
• 金额: $ 15万
• 依托单位: Fyodor Biotechnologies, Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1215242&HistoricalAwards=false
• 关键词: SBIR; Phase; Recombinant; Multiepitope; Mosaic

This Small Business Innovation Research (SBIR) Phase I project aims to develop recombinant monoclonal antibodies using "mosaic" antigens derived from Leptospira spp proteins identified in the urine of clinical patients. The mosaic antigens are designed to artificially combine important antigenic epitopes in order to maximize the coverage of potential epitopes in one structure and to provide broader coverage than those afforded by multiple synthetic peptides. Our approach involves the use of computational biology and bioinformatics to create, score, and select "mosaic" antigens from Leptospira spp. Antigenic properties of the mosaic antigens are evaluated by indirect ELISA using a panel of well-characterized human sera from clinical patients and apparently healthy individuals. We will then use recombinant DNA and protein engineering techniques to derive cognate chimeric proteins, followed by the generation of recombinant epitope-specific monoclonal antibodies that target potential epitopes (rather than a few dominant epitopes) generated from the "mosaic" protein. The diagnostic potential of the recombinant antibody is evaluated by immunoblotting and ELISA using leptospirosis specimens from early and convalescent phases of the illness.The broader impact/commercial potential of this project, if successful, will be the development of a rapid, non-invasive, one-step, multiplex test that will have general applicability in the timely diagnosis of a complement of febrile illnesses, including leptospirosis. This is aimed at tropical and subtropical countries where the disease incidences are higher and where current tools are mostly inadequate. Early leptospira diagnosis is essential because antibiotic treatment is most effective when initiated early in the course of the disease. Current diagnostic tools are unsuitable for use in resource-limited settings especially in the tropical regions where other similar acute febrile illnesses are common. It expands the multiplex diagnostic test panel we are presently developing by iterative expansion.

这项小企业创新研究（SBIR）一期项目旨在利用从临床患者尿液中鉴定的钩端螺旋体蛋白衍生的“镶嵌”抗原开发重组单克隆抗体。马赛克抗原被设计为人工组合重要的抗原表位，以最大限度地在一个结构中覆盖潜在的表位，并提供比多个合成肽更广泛的覆盖范围。我们的方法包括使用计算生物学和生物信息学来从钩端螺旋体中创建、评分和选择“马赛克”抗原。使用一组来自临床患者和表面健康个体的特征良好的人血清，通过间接ELISA评估马赛克抗原的抗原特性。然后，我们将使用重组DNA和蛋白质工程技术获得同源嵌合蛋白，随后生成重组表位特异性单克隆抗体，该抗体针对从“马赛克”蛋白产生的潜在表位（而不是少数显性表位）。利用钩端螺旋体病早期和恢复期的标本，通过免疫印迹法和ELISA法评估重组抗体的诊断潜力。如果该项目成功，其更广泛的影响/商业潜力将是开发一种快速、无创、一步、多重测试，这种测试将在及时诊断包括钩端螺旋体病在内的一系列发热性疾病方面具有普遍适用性。这是针对热带和亚热带国家的，在这些国家，疾病发病率较高，目前的工具大多不足。钩端螺旋体的早期诊断是必不可少的，因为在疾病的早期开始抗生素治疗是最有效的。目前的诊断工具不适合在资源有限的环境中使用，特别是在其他类似急性发热性疾病常见的热带地区。它扩展了我们目前通过迭代扩展开发的多路诊断测试面板。